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. 1998 Sep 29;95(20):11566–11571. doi: 10.1073/pnas.95.20.11566

Figure 2.

Figure 2

Cleavage of PKN by caspase-3-like protease. (A) Inhibition of PKN cleavage during apoptosis by DEVD-CHO. Jurkat cells were pretreated with various concentrations of the tetrapeptide inhibitor of caspase-1 (YVAD-CHO) or caspase-3 (DEVD-CHO) for 1 h, as indicated, and then incubated with α-Fas mAb (150 ng/ml) for an additional 6 h. The cleavage of PKN was analyzed by immunoblotting with αC6. (B) In vitro cleavage of PKN by recombinant caspases. His-PKN (40 ng) was incubated with recombinant His-caspase-3 (Casp-3) or GST-caspase-6 (Casp-6) at 30°C for the indicated time periods, then analyzed by immunoblotting with PKN (C-19). The cleavage products corresponding to AF1, AF2, and AF3 were observed from 10-min incubation with caspase-3, while a cleavage product of a different size was observed from caspase-6 cleavage. White and black arrowheads on the right of the blot indicate the proteins uncleaved and cleaved by caspase-3, respectively. Gray arrowhead indicates a caspase-3 cleavage product that was not detected in vivo. Molecular mass markers in kDa are indicated on the left.